Automated liquid manufacturing system

ABSTRACT

A method for continuously preparing a medium formulation mixes a diluent with a plurality of chemically incompatible concentrate solutions in such a manner that none of the ingredients of the concentrate solutions chemically react in an adverse manner. The method utilizes a static mixing chamber to add the concentrate solutions to the diluent stream sufficiently in advance of one another so that adverse chemical reactions do not occur. The method also adjusts a pH level of the diluent prior to adding any of the concentrate solutions to the diluent.

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] The present invention relates to the field of cell culture mediumformulations, and more specifically, to methods for continuouslypreparing cell culture medium formulations and buffered salt solutionsfrom selected subgroups of medium concentrates.

[0003] 2. Background Art

[0004] Cell culture medium formulation provide nutrients necessary tomaintain and grow cells in a controlled, artificial and in vitroenvironment. Characteristics and compositions of the cell culturemediums vary depending on the particular cellular requirements.Important parameters include osmolarity, pH, and nutrient formulations.

[0005] Medium formulations have been used to grow a number of cell typesincluding animal, plant and bacterial cells. Cells grown in culturemedium catabolize available nutrients and produce useful biologicalsubstances such as monoclonal antibodies, hormones, growth factors andthe like. Such products have therapeutic applications and, with theadvent of recombinant DNA technology, cells can be engineered to producelarge quantities of these products. Thus, the ability to grow cells invitro is not only important for the study of cell physiology, it isnecessary for the production of useful substances which may nototherwise be obtained by cost-effective means.

[0006] Cell culture medium formulations have been well documented in theliterature and a number of medium are commercially available. Typicalnutrients in cell culture medium formulations include amino acids,salts, vitamins, trace metals, sugars, lipids and nucleic acids. Often,particularly in complex medium formulations, stability problems resultin toxic products and/or lower effective concentrations of requirednutrients, thereby limiting the functional life-span of the culturemedium. For instance, glutamine is a constituent of almost all mediumformulations that are used in the culturing of mammalian cells in vitro.Glutamine decomposes spontaneously into pyrrolidone carboxylic acid andammonia. The rate of degradation can be influenced by pH and ionicconditions but in cell culture medium, formation of these breakdownproducts cannot be avoided (Tritsch et al., Exp. Cell Research,28:360-364(1962)).

[0007] Wang et al. (In Vitro, 14:(8):715-722 (1978)) have shown thatphotoproducts such as hydrogen peroxide, which are lethal to cells, areproduced in Dulbecco's Modified Eagle's Medium (DMEM). Riboflavin andtryptophan or tyrosine are components necessary for formation ofhydrogen peroxide during light exposure. Because most mammalian culturemedium formulations contain riboflavin, tyrosine and tryptophan, toxicphotoproducts are likely produced in most cell culture mediums.

[0008] To avoid these problems, researchers make medium formulations onan “as needed” basis, and avoid long term storage of the culture medium.Commercially available medium formulations, typically in dry powderform, serve as a convenient alternative to making the mediumformulations from scratch, i.e., adding each nutrient individually, andalso avoids some of the stability problems associated with liquid mediumformulations. However, only a limited number of commercial culturemedium formulations are available, except for those custom formulationssupplied by the manufacturer.

[0009] Although dry powder medium formulations may increase theshelf-life of some medium formulations, there are a number of problemsassociated with dry powdered medium formulations, especially in largescale application. Production of large volumes requires storagefacilities for the dry powder, not to mention the specialized kitchensnecessary to mix and weigh the nutrient components. Due to the corrosivenature of dry powder medium ingredients, mixing tanks must beperiodically replaced.

[0010] There exists a need to lower the cost of production of biologicalsubstances. Efficient and cost effective methods to stabilize liquidcell culture medium formulations as well as the development ofconvenient methods to produce 1× medium formulations would be animportant development in the field of cell culture medium technology.

[0011] One such development in the field of cell culture mediumformulations is the development of liquid medium concentrates as isdisclosed in U.S. Pat. No. 5,474,931 issued to DiSorbo et al. on Dec.12, 1995 (“DiSorbo”). DiSorbo discloses a method of subgrouping mediumformulations into stable, compatible components that can be solubilizedat high concentrations (10× to 100×). Concentrated culture mediumformulations (2-10×) or 1× cell culture medium formulations can beprepared by mixing a sufficient amount of the concentrated subgroupsolutions with each other and with a sufficient amount of a diluent(water, buffer, etc.).

[0012] Escalating demand for large volumes of nutrient medium andbuffered salt solutions and increasing pressure to minimizebatch-associated costs, such as sterile filtration and quality releasetesting, has driven a requirement for increased production batch sizesof liquid medium. As a result, stainless steel formulation tanks of5000-10,000 liters for preparation of large batches of liquid medium orbuffered salt solutions have become relatively common. However, scale-upmanufacture of these fluids in this manner presents challenges regardingproduct quality and economy.

[0013] What is needed is a system and method for providing continuous,online preparation of large volumes of biological fluids (e.g., liquidmedium, buffered salt solutions, etc.) within a highly controlledmanufacturing system.

BRIEF SUMMARY OF THE INVENTION

[0014] The present invention is a system and method for continuous,online preparation of cell culture medium formulations from selectedsubgroups of medium concentrates. In particular, a computer controlledsystem controls the flow of a diluent and one or more concentratedsolutions into a static mixing chamber wherein the diluent and theconcentrated solutions are mixed to form the cell culture mediumformulations.

[0015] The present invention is able to formulate a cell culture mediumfrom concentrated solution subgroups including an acid solubleconcentrate solution subgroup, a group I salts solution concentratesubgroup, a group II salts solution concentrate subgroup, and a basesoluble solution concentrate subgroup. Furthermore, the presentinvention is able to adjust the pH of the cell culture medium usingeither an acid solution or a base (caustic) solution.

[0016] In particular, the present invention is able to mix theconcentrated solution subgroups with the diluent in a manner such thatthe ingredients of the concentrated solution subgroups do not adverselyreact chemically with one another.

[0017] One feature of the present invention is the preparation of largequantities of 1× cell culture medium (100,000 liters or more) whilerequiring only one quality control test. By increasing the size of the“batch,” the present invention reduces the per liter cost of cellculture medium.

[0018] Another feature of the present invention is the increasedconsistency in the 1× cell culture medium. Statistical analyses havedemonstrated that the present invention is able to provide 1× cellculture medium with homogeneity within batches of ±2.0%. Furthermore,the present invention provides improved precision between productionruns of 1× cell culture medium manufactured from identical concentratesolutions of ±3.0%.

[0019] Still another feature of the present invention is a clean inplace (CIP) and a steam in place (SIP) system which allows variouscomponents of the present invention to be sanitized and sterilizedaccording to current good manufacturing practices (cGMP).

[0020] Further features and advantages of the present invention, as wellas the structure and operation of various embodiments of the presentinvention, are described in detail below with reference to theaccompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

[0021] The present invention is described with reference to theaccompanying drawings. In the drawings, like reference numbers indicateidentical or functionally similar elements. Additionally, the left-mostdigit(s) of a reference number identifies the drawing in which thereference number first appears.

[0022]FIG. 1 illustrates an automated liquid manufacturing system (ALMS)according to the present invention.

[0023]FIG. 2 illustrates a diluent system according to a preferredembodiment of the present invention.

[0024]FIG. 3 illustrates a medium mixing system according to a preferredembodiment of the present invention.

[0025]FIG. 4 illustrates a medium surge vessel according to oneembodiment of the present invention.

[0026]FIG. 5 illustrates a pre-filtration system and a sterilefiltration system according to a preferred embodiment of the presentinvention.

[0027]FIG. 6A and 6B, respectively, illustrate a front view and a rightside view of a medium mixing chamber according to a preferred embodimentof the present invention.

[0028]FIG. 7 illustrates an isometric view of a portion of the mediummixing chamber according to a preferred embodiment of the presentinvention.

[0029]FIG. 8 illustrates an example of a computer control system usefulfor controlling the operation of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

[0030] In the description that follows, a number of terms conventionallyused in the field of cell culture medium are utilized extensively. Inorder to provide a clear and consistent understanding of thespecification and claims, and the scope to be given such terms, thefollowing definitions are provided.

[0031] Ingredients. The term “ingredients” refers to any compound,whether of chemical or biological origin, that can be used in cellculture medium to maintain or promote the growth or proliferation ofcells. The terms “component,” “nutrient,” and “ingredient” can be usedinterchangeably and are all meant to refer to such compounds. Typicalingredients that are used in cell culture medium formulations includeamino acids, salts, metals, sugars, lipids, nucleic acids, hormones,vitamins, fatty acids, proteins and the like. Other ingredients thatpromote or maintain growth of cells in vitro can be selected by those ofskill in the art, in accordance with the particular need.

[0032] Cell Culture. By “cell culture” is meant cells or tissues thatare maintained, cultured or grown in an artificial, in vitroenvironment.

[0033] Culture Vessel. Glass, plastic or metal containers of varioussizes that can provide an aseptic environment for growing cells aretermed “culture vessels.”

[0034] Cell Culture Medium. The phrases “cell culture medium” or“culture medium” or “medium formulation” or “cell culture mediumformulation” refer to a nutritive solution for culturing or growingcells. The ingredients that comprise such medium formulations may varydepending on the type of cell to be cultured. In addition to nutrientcomposition, osmolarity and pH are considered important parameters ofculture medium formulations.

[0035] Compatible Ingredients. Each ingredient used in cell culturemedium formulations has unique physical and chemical characteristics. By“compatible ingredients” is meant those medium nutrients which can bemaintained in solution and form a “stable” combination. A solutioncontaining “compatible ingredients” is said to be “stable” when theingredients do not degrade or decompose substantially into toxiccompounds, or do not degrade or decompose substantially into compoundsthat can not be utilized or catabolized by the cell culture. Ingredientsare also considered “stable” if degradation can not be detected or whendegradation occurs at a slower rate when compared to decomposition ofthe same ingredient in a 1× cell culture medium formulation. Glutamine,for example, in 1× medium formulations, is known to degrade intopyrrolidone carboxylic acid and ammonia. Glutamine in combination withdivalent cations are considered “compatible ingredients” since little orno decomposition can be detected over time.

[0036] Compatibility of medium ingredients, in addition to stabilitymeasurements, are also determined by the “solubility” of the ingredientsin solution. The term “solubility” or “soluble” refers to the ability ofan ingredient to form a solution with other ingredients. Ingredients arethus compatible if they can be maintained in solution without forming ameasurable or detectable precipitate. Thus, the term “compatibleingredients” as used herein refers to the combination of particularculture medium ingredients which, when mixed in solution either asconcentrated or 1× medium formulations, are “stable” and “soluble.”

[0037] 1× Formulation. A cell culture medium is composed of a number ofingredients and these ingredients vary from medium to medium. A “1×formulation” or “1× medium formulation” is meant to refer to any aqueoussolution that contains some or all ingredients found in a cell culturemedium. The “1× formulation” can refer to, for example, the cell culturemedium or to any subgroup of ingredients for that medium. Theconcentration of an ingredient in a 1× solution is about the same as theconcentration of that ingredient found in the cell culture formulationused for maintaining or growing cells. Cell culture medium formulationsused to grow cells are 1× formulation by definition. When a number ofingredients are present (as in a subgroup of compatible ingredients),each ingredient in a 1× formulation has a concentration about equal tothe concentration of those ingredients in a cell culture medium. Forexample, RPMI 1640 culture medium contains, among other ingredients, 0.2g/l L-arginine, 0.05 g/l L-asparagine, and 0.02 g/l L-aspartic acid. A“1× formulation” of these amino acids, which are compatible ingredientsaccording to the present invention, contains about the sameconcentrations of these ingredients in solution. Thus, when referring toa “1× formulation,” it is intended that each ingredient in solution hasthe same or about the same concentration as that found in the cellculture medium being described. The concentrations of medium ingredientsin a 1× formulation are well known to those of ordinary skill in theart, See Methods For Preparation of Media, Supplements and Substrate ForSerum-Free Animal Cell Culture, Allen R. Liss, N.Y. (1984), which isincorporated by reference herein in its entirety. The osmolarity and/orpH, however, may differ in a 1× formulation compared to the culturemedium, particularly when fewer ingredients are contained by the 1×formulation.

[0038] 10× Formulation. A “10× formulation” refers to a solution whereineach ingredient in that solution is about 10 times more concentratedthan the same ingredient in the cell culture medium formulation. RPMI1640 medium, for example, contains, among other things, 0.3 g/lL-glutamine. By definition, a “10× formulation” contains about 3.0 g/lglutamine. A “10× formulation” may contain a number of additionalingredients at a concentration about 10 times that found in the 1×culture medium. As will be apparent, “25× formulation,” “50×formulation” and “100× formulation” designate solutions that containingredients at about 25, 50 or 100 fold concentrations, respectively, ascompared to a 1× cell culture medium. Again, the osmolarity and pH ofthe medium formulation and concentrated formulation may vary.

[0039] Automated Liquid Manufacturing System

[0040] According to the present invention, an automated liquidmanufacturing system (ALMS) continuously prepares medium products (e.g.,cell culture medium, buffered salt solutions, salt solutions, buffers,etc.) having various formulations (e.g., 1-10×) by mixing one or moreconcentrate solution subgroups together with a diluent (e.g. water,buffer, etc.). The amount of concentrated solution and amount of diluentneeded may vary depending on the concentration of each subgroup, thenumber of subgroups, and the desired concentration of the final mediumproduct. One of ordinary skill in the art can easily determined asufficient volume of a diluent and a sufficient volume of theconcentrated solutions to prepare the desired medium product.

[0041] The pH of the desired medium product may also be adjusted by theaddition of acid or base. The medium product, however, may not requireany adjustment, especially if the pH of the medium product as preparedis within the desired pH range. Osmolarity of the medium product canalso be adjusted after mixing the concentrated solutions with thediluent. Typically, the desired osmolarity may be predetermined andadjustments in the salt concentration of the concentrated solutions maybe made to prepare a final medium product with the desired osmolarity.

[0042] The present invention also provides for on-line sanitization andsterilization in place as required by current good manufacturingpractices (cGMP). The sanitization operation is commonly referred to as“clean in place,” and sterilization operation is commonly referred to as“steam in place.” These operations are discussed in further detailbelow.

[0043] According to the present invention, sufficient amounts of eachconcentrate solution subgroup are continuously admixed with sufficientamounts of a diluent in a mixing chamber, while the resulting mediumproduct is continuously removed. The following describes various aspectsof the present invention and the manner in which they accomplish thecontinuous preparation of medium product.

[0044]FIG. 1 illustrates a system level block diagram of an automatedliquid manufacturing system (ALMS) 100 according to the presentinvention. ALMS 100 includes a concentrate system 110, a diluent system120, a medium mixing system 130, a medium surge vessel 140, aprefiltration system 150, a sterile filtration system 160 and a fillsystem 170. Sterile filtration system 160 and fill system 170 operate ina clean area 180. In addition to the above-mentioned system components,a preferred embodiment of the present invention includes a wastedisposal system 190. Each of these components of ALMS 100 will bediscussed in further detail below.

[0045] A preferred embodiment of the present invention is controlled bya computer control system 105. For ease of illustration, connectionsbetween computer control system 105 and the various components of ALMS100 have not been shown. Needless to say, each of the components of ALMS100 has some subcomponent, be it a valve, a pump, a sensor, etc., thatis connected to computer control system 105 and used to control theoperation of ALMS 100 as would be apparent. Computer control system 105is described in further detail below.

[0046] Concentrate System

[0047] Concentrate system 110 provides one or more concentrate solutions115 to ALMS 100. Specifically, concentrate system 110 providesconcentrate solutions 115 to medium mixing system 130. Concentratesystem 110 may perform this task in a variety of ways. In one embodimentof the present invention, concentrate system may provide concentratesolutions 115 in a manner similar to that described in commonly ownedU.S. Pat. No. 5,474,931 issued to DiSorbo et al. on Dec. 12, 1995, whichis incorporated herein by reference as if reproduced below in itsentirety. DiSorbo discloses a method for producing liquid mediumconcentrates in compatible subgroups. According to this embodiment ofthe present invention, concentrate solutions 115 are chemically stable50× formulations of liquid medium concentrates.

[0048] These subgroups include the following: an acid solubleconcentrate solution subgroup, a group I salts concentrate solutionsubgroup, a group II concentrate solution subgroup, and a base solubleconcentrate solution subgroup.

[0049] In addition, sodium hydroxide may be prepared as a concentratesolution subgroup although this is not necessary. The acid solubleconcentrate solution subgroup referred to herein is essentiallyequivalent to the acid-soluble subgroup referred to in DiSorbo; thegroup I salts concentrate solution subgroup referred to herein isessentially equivalent to the glutamine-containing subgroup referred toin DiSorbo; the group II salts concentrate solution subgroup referred toherein is essentially equivalent to the weak acid-base soluble subgroupreferred to in DiSorbo; and the base soluble concentrate solutionsubgroup referred to herein is essentially equivalent to thealkali-soluble subgroup referred to in DiSorbo. The remaining subgroupsreferred to in DiSorbo are treated as reserve concentrate solutions forpurposes of the present invention.

[0050] In this embodiment, the subgroups are formulated and “kited”according to published procedures as would be apparent. After beingprepared according to these procedures the subgroups are stored inintermediate storage vessels for use by ALMS 100.

[0051] In another embodiment of the present invention, concentratesystem 110 provides preformulated and prepackaged concentrate solutions115. These concentrate solutions 115 are purchased from a manufacturerof such concentrate solutions such as are available from LifeTechnologies, Incorporated, 3175 Staley Road, Grand Island, N.Y.,716/774-6700. In addition, concentrated subgroups for buffered salts canbe obtained from Life Technologies as acid soluble concentrate solutionsubgroups and base soluble concentrate solution subgroups. Thisembodiment permits a manufacturer of medium products to purchaseconcentrate solutions 115 without itself having the facilities tomanufacture or produce such concentrate solutions 115.

[0052] In yet another embodiment of the present invention, concentratesystem 110 provides an on-line concentrate solution 115 as a part of acontinuous manufacturing process in which concentrate solutions 115 areproduced directly from raw materials and passed directly to ALMS 100without an intermediate storage device such as that described inDiSorbo.

[0053] As would be apparent to one skilled in the art, other types ofconcentrate solutions 115 are available other than the subgroupsdescribed above. Furthermore, other means for providing concentratesolution 115 to ALMS 100 may be available as would also be apparent.

[0054] Diluent System

[0055] Diluent system 120 provides a diluent 125 to ALMS 100. Inparticular, diluent system 120 provides diluent 125 to medium mixingsystem 130. Diluent 125 may be any solution or liquid that may be usedto dilute concentrate solutions 115. Such diluents include water,buffers, salt solutions, etc. In a preferred embodiment of the presentinvention, diluent 125 is water, most preferably, water for injection.However, any diluent 125 may be used in ALMS 100 that appropriatelydilutes concentrate solutions 115 according to the particular needs ofthe medium product manufacturer.

[0056] A preferred embodiment of diluent system 120 is illustrated inFIG. 2. In this embodiment of the present invention, diluent system 120includes an ambient water for injection (WFI) tank 210, a hot WFI tank220, a control valve 215, a control valve 225, and a WFI break tank 230.WFI break tank 230 includes a level indicator 250 and a spray ball 240.

[0057] The purpose of WFI break tank 230 is to provide an atmosphericbreak between the plant water system and ALMS 100 as required by currentgood manufacturing practices (cGMP). In addition, WFI break tank 230assures removal of entrained air from ambient WFI tank 210 and hot WFItank 220 prior to their introduction to ALMS 100.

[0058] In one embodiment of the present invention, ambient WFI tank 210is not a tank. Rather, ambient WFI tank 210 is directly connected to theplant's water system. In other embodiments of the present invention,ambient WFI tank 210 may actually be a tank. This may be the case, forexample, when a diluent 125 other than water is used, or when aparticular type of water is required (e.g. deionized, distilled,sterile, etc.). Hot WFI tank 220 provides hot water to ALMS 100 during aclean-in-place (CIP) operation which is discussed in further detailbelow.

[0059] Valve 215 and valve 225 control the flow of ambient water fromambient WFI tank 210 and hot water from hot WFI tank 220, respectively,to WFI break tank 230. In a preferred embodiment of the presentinvention, WFI break tank 230 provides ambient water as diluent 125 toALMS 100.

[0060] Level indicator 250 monitors a level of diluent 125 in WFI breaktank 230. Level indicator 250 is monitored by computer control system105 to maintain an appropriate level of diluent 125 in WFI break tank230.

[0061] Spray ball 240 is a part of the CIP operation which is discussedin further detail below. Spray ball 240 provides a mechanism forcleaning the inside of WFI break tank 230 during the CIP operation.

[0062] Medium Mixing System

[0063] Medium mixing system 130 is shown in further detail in FIG. 3.Medium mixing system 130 includes a static mixing chamber 310, a diluentinput pump 320, a diluent flow indicator 325, a CIP divert valve 330, aseries of concentrate solution pumps 340 (shown as concentrate solutionpumps 340A-H), a first pH sensor 361, a second pH sensor 362, aconductivity sensor 363, a UV absorbance sensor 364, an output flowindicator 365, a diverter valve 370, and a back flow preventer valve375. Each of these elements of medium mixing system 130 is described infurther detail below.

[0064] Medium mixing system 130 receives diluent 125 and one or moreconcentrate solutions 115 and mixes them in mixing chamber 310. Mediummixing system 130 accomplishes this in a manner such that none of theingredients of concentrate solutions 115 adversely chemically react withone another or with diluent 125. By “adversely chemically react” it ismeant that the ingredients react 1) to form an irreversible precipitate;2) to cause degradation in one or more components of the concentratesolutions; 3) to cause certain components to become inactivated; or 4)to cause any other condition that would result in an unacceptable mediumproduct 135.

[0065] Diluent input pump 320 controls the flow of diluent 125 intostatic mixing chamber 310. This flow is measured by diluent flowindicator 325. Diluent flow indicator 325 permits computer controlsystem 105 to monitor the flow of diluent 125 and thereby, controldiluent input pump 320. Back flow preventer valve 375 prevents diluent125 from flowing backwards from static mixing chamber

[0066] Based on the flow of diluent 125 into static mixing chamber 310,computer control system 105 controls the flows of concentrate solutions115 (shown as concentrate solutions 115A-H) into static mixing chamber310 via concentrate solution pumps 340 (shown as concentrate solutionpumps 340A-H). The flow of each of concentrate solutions 115A-H iscontrolled to be proportional to the flow of diluent 125 into staticmixing chamber 310 according to a formulation of a desired mediumproduct.

[0067] Sensors 361, 362, 363, 364 and 365 monitor a medium product 135output from static mixing chamber 310 to ensure that particularparameters associated with medium product 135 are within acceptablelevels associated with the desired medium product. These sensors arecoupled to computer control system 105 which monitors these parametersof medium product 135 to ensure that proper mixing of concentratesolutions 115A-H and diluent 125 is being accomplished. If the mediumproduct is within the acceptance levels, medium product 135 passes tomedium surge vessel 140. If not, computer control system 105 divertsmedium product 135 to waste disposal system 190 via diverter valve 370.This allows medium mixing system 130 to guarantee an acceptable mediumproduct 135. For example, when ALMS 100 starts up preparation of aparticular medium product 135, the initial output of static mixingchamber 310 may not be within the acceptance levels for the particularmedium product. Thus, this portion of the output is diverted to wastedisposal system 190. When the output of static mixing chamber 310 entersinto the acceptable levels (i.e., the operation reaches a “steadystate”), the output from static mixing chamber 310 is passed to mediumsurge vessel 140.

[0068] In a preferred embodiment of the present invention, first pHsensor 361 and second pH sensor 362 are placed in close proximity toeach other and as close to static mixing chamber 310 as possible, andprior to sensors 363, 364 to ensure that the proper pH levels of mediumproduct 135 is being achieved.

[0069] Conductivity sensor 363 measures the ionic character of mediumproduct 135. In particular, conductivity sensor 363 measures theresistivity of the flow of medium product 135. Conductivity sensor 363is useful for determining the quality of medium product 135, especiallyfor salt solutions.

[0070] UV absorbance sensor 364 measures an amount of ultraviolet lightthat passes through the flow of medium product 135. UV absorbance sensor364 is useful for detecting the presence of precipitates within mediumproduct 135. UV absorbance sensor 364 can also be used to measure aconcentration of a particular component as an on-line measurement ofconcentrate addition and mixing quality.

[0071] As would be apparent to one skilled in the art, other types ofsensors may be implemented in medium mixing system 130 to measure otherlevels of other parameters associated with medium product 135.

[0072] In a preferred embodiment of the present invention, concentratesolution pumps 340A-H are extremely precise variable speed pumps. Inparticular, concentrate solution pumps 340A-F are capable of delivering0 to 3 liters of fluid per minute with ±1.0% or better accuracy.Concentrate solution pumps 340G-H are capable of delivering 0 to 3.5liters of fluid per minute with ±1.0% accuracy. A preferred embodimentof the present invention uses pumps which are manufactured by IVEK,North Springfield, Vt.

[0073] In a preferred embodiment of the present invention, concentratesolution 115A and concentrate solution 115B are reserved for providingan acid solution and a base solution, respectively, to static mixingchamber 310. Hence, referring to these as “concentrate solutions” may beconsidered a misnomer. However, as would be apparent, solutions,liquids, etc., other that “concentrate solutions” may be introduced inthis manner to static mixing chamber 310 as would be apparent.

[0074] In this preferred embodiment of the present invention, acidsolution 115A and caustic solution 115B adjust a pH level of diluent 125according to specifications required by the production of medium product135. The addition of either acid solution 115A or caustic solution 115Bto diluent 125 is done first so that the proper pH level of diluent 125can be achieved prior to the addition of other concentrate solutions115C-H.

[0075] As shown in FIG. 3, diluent 125 enters static mixing chamber 310and begins “mixing” sufficiently prior to the addition of anyconcentrate solutions 115A-H. This ensures that static mixing chamber310 can provide a “turbulent diluent stream” from diluent 125 to enhancethe overall mixing process between diluent steam 125 and concentratesolution 115A-H. The turbulent diluent stream is produced from diluent125 by being forced past a series of baffles within static mixingchamber 310 as is well understood by those in the art. Also, theintroduction of a last concentrate solution 115H occurs sufficientlyprior to the end of static mixing chamber 310 so that last concentratesolution 115H can be sufficiently mixed in turbulent diluent stream. Asdiscussed above, the output of static mixing chamber 310 is mediumproduct 135.

[0076] As shown in FIG. 3, static mixing chamber 310 includes a seriesof injection ports 315 (shown as injection ports 315A-315H). Injectionports 315 introduce concentrate solutions 115 into static mixing chamber310. In particular, injection ports 315 introduce concentrate solutions115 into turbulent diluent stream 125. FIG. 6 shows a mechanical drawingof static mixing chamber 310 in further detail.

[0077]FIG. 6A, FIG. 6B, and FIG. 7 illustrate static mixing chamber 310in greater detail. In particular, FIGS. 6A and 6B are mechanicaldrawings showing a front view and a right side view, respectively, ofstatic mixing chamber 310. FIG. 7 is an isometric drawing of staticmixing chamber 310. As shown in FIGS. 6A, 6B, and 7, static mixingchamber 310 includes a series of injection ports 315. In particular,static mixing chamber 310 includes two groupings of radially disposedinjection ports shown as injection ports 315C, 315D, and 315E andinjection ports 315F, 315G, and 315H. In addition, as shown in FIGS. 6Aand 6B, static mixing chamber 310 also includes two additional injectionports 315A and 315B.

[0078] Injection ports 315C, 315D, and 315E are described as beingradially disposed around static mixing chamber 310. By “radiallydisposed” it is meant that injection ports 315C, 315D, and 315E arelocated on a common circumference around static mixing chamber 310. Thatis, injection ports 315C, 315D, and 315E are located at an approximatelyequal distance from the upstream end of static mixing chamber 310.Preferably, injection ports 315C, 315D, and 315E are spaced equallyabout the common circumference of static mixing chamber 310. Thus, forthe case of three injection ports, the injection ports 315C, 315D, and315E are space at 120 degree increments. Other embodiments may providefor non-equal spacings about the common circumference.

[0079] In one embodiment of the present invention, the injection portsare essentially disposed both “linearly” and “radially” from oneanother. Such would be the case, for example, where the injection portswere disposed in spiral fashion about static mixing chamber 310.Depending on the length of the spiral, the injection ports could beconsidered linearly disposed, radially disposed, or both. Injectionports 315F, 315G, and 315H are also radially disposed around staticmixing chamber 310. In addition, this group of injection ports, bothindividually and collectively, is “linearly disposed” along the fluidflow path of static mixing chamber 310 from injection ports 315C, 315D,and 315E as shown in FIG. 6. In other words, injection ports 315F, 315G,and 315H are located at an approximately equal distance from theupstream end of static mixing chamber 310, where this distance issufficiently different from the distance from the upstream end of staticmixing chamber 310 to injection ports 315C, 315D, and 315E.

[0080] In the particular embodiment shown in FIG. 6 and FIG. 7, threeinjection ports are radially disposed from one another in each of thetwo groups of injection ports. As would be apparent to one skilled inthe art, additional injection ports may be included within each group,limited by two parameters. The first parameter is the number ofinjection ports that can physically, or mechanically, fit around staticmixing chamber 310. The second parameter is the number of injectionports that can be used to introduce concentrate solutions 115 to diluent125 without the ingredients of concentrate solutions 115 adverselychemically reacting with one another. As also would be apparent, fewerinjection ports may be included within each group.

[0081] In addition to changing the number of injection ports within eachradially disposed group, the number of radially disposed groups may alsobe changed. The number of radially disposed groups of injection ports isalso limited by the same parameters as described above as would beapparent.

[0082] As shown in FIG. 6 and FIG. 7, diluent 125 flows from theupstream end of static mixing chamber 310 toward the downstream end ofstatic mixing chamber 310. Thus, as diluent 125 flows through staticmixing chamber 310, diluent 125 encounters injection ports 315A and 315Bfirst, followed by injection ports 315C, 315D and 315E, and finally,injection ports 315F, 315G and 315H.

[0083] As thus described, static mixing chamber 310 provides two mannersin which different concentrate solutions 115 can be added to diluent125. The first manner is to add the different concentrate solutions 115by using injection ports that are radially disposed from one anothersuch as injection ports 315F, 315G, 315H or injection ports 315C, 315Dand 315E. The second manner in which different concentrate solutions 115can be added to diluent 125 is by using injection ports 315 that arelinearly disposed from one another such as injection ports 315C and315F. In either case, an injection port 315 adds a concentrate solution115 to diluent 125 in a manner such that the concentrate solution 115becomes sufficiently diluted by diluent 125 prior to encountering anyother concentrate solution 115 added from a different injection port315. This prevents any adverse chemical reaction between the ingredientsof the two concentrate solutions.

[0084] While this is true in general, the order of introduction ofcertain concentrate solutions 115 to diluent 125 from a particularinjection port configuration are preferred, while other orders ofintroduction are discouraged. For example, medium product 135 thatincludes a base soluble concentrate solution and a group II saltsconcentrate solution are preferably prepared by introducing these twoconcentrate solutions into diluent 125 by radially disposed injectionports. Doing so improves the microenvironment chemistry of the resultingmedium product 135.

[0085] Also, medium product 135 that includes a group II saltsconcentrate solution and an acid soluble concentrate solution arepreferably prepared by introducing these two concentrate solutions intodiluent 125 from linearly disposed injection ports 315. Introducingthese two concentrate solutions from injection ports that are radiallydisposed from one another is detrimental to product quality and maycreate an irreversible precipitation of critical cell culture mediumcomponents rendering the resulting medium product inactive.

[0086] In a preferred embodiment of the present invention, the followinginjection ports 315 concentrate solution 115 pairings are used: acidsoluble concentrate solutions are introduced by injection port 315D;group I salts concentrate solutions are introduced by injection port315E; group II salts concentrate solutions are introduced by injectionport 315G; base soluble concentrate solutions are introduced byinjection port 315H; acid solutions for adjusting pH are introduced byinjection port 315A; and base (caustic) solutions for adjusting pH areintroduced by injection port 315B. If sodium hydroxide concentratesolutions are used, they are preferably introduced by injection port315F. Otherwise, injection port 315F is reserved for other concentratesolutions not included above. Injection port 315C is also reserved forother concentrate solutions not included above.

[0087] Medium Surge Vessel

[0088]FIG. 4 illustrates medium surge vessel 140 in greater detail.Medium surge vessel 140 includes a medium surge tank 410, an agitationsystem 420, a level indicator 430, a temperature control system 450, anda pH sensor 470. Medium product 135 from medium mixing system 130 entersmedium surge tank 410 which provides a buffering mechanism for ALMS 100.In other words, medium surge vessel 140 provides a “buffer” between thecontinuous operation of medium mixing system 130 and the discontinuousoperation of downstream components of ALMS 100 such as fill system 170.Thus, medium product 135 from medium mixing system is permitted toaccumulate in medium surge vessel 140 when, for example, fill system 170is temporarily shutdown to change fill containers.

[0089] An amount of medium product 135 in medium surge tank 410 ismonitored by computer control system 105 via fill indicator 430.Depending on the level of medium product 135 in medium surge tank 410,computer control system 105 adjusts the output rate of medium product135 from medium mixing system 130.

[0090] A pH level of medium product 135 is measured by pH sensor 470 asmedium product 135 leaves medium surge tank 410. This permits computercontrol system 105 to monitor and ensure the quality of medium product135.

[0091] In one embodiment of the present invention, agitation system 420is used to provide agitation (i.e., mixing) to medium product 135 withinmedium surge tank 410. In one embodiment, agitation system 420 providescontinuous mixing of medium product 135 in medium surge tank 410. Inanother embodiment, agitation system 420 provides mixing of mediumproduct in medium surge tank 410 after a particular level is reached orsome other parameter. Agitation system 420 may or may not be required inorder to maintain medium product 135 in a homogeneous state. In apreferred embodiment of the present invention, agitation system 420 isnot used.

[0092] In one embodiment of the present invention, temperature controlsystem 450 controls the temperature of medium product 135 within mediumsurge tank 410. Temperature control system 450 operates so as tomaintain a particular temperature of medium product 135 in medium surgetank 410. Various means of controlling the temperature of the contentsof medium surge tank 410 are available as would be apparent. In oneembodiment of the present invention, glycol is circulated through anouter tank (not shown) around medium surge tank 410 thereby maintaininga particular temperature of the contents within medium surge tank 410.In a preferred embodiment of the present invention, temperature controlsystem 450 is not used.

[0093] In one embodiment of the present invention, compressed air 460 isprovided to medium surge tank 410 to maintain a given head pressurewithin medium surge tank 410. Compressed air 460 is used to providesufficient pressure to move medium product 135 through medium surge tankinto prefiltration system 150. In a preferred embodiment of the presentinvention, the head pressure is maintained between 6 and 10 p.s.i.g.Other embodiments may utilize gases other than air, such as nitrogen, toprovide the head pressure as well as to prevent the outgasing frommedium product 135 as would be apparent.

[0094] Diverter valve 445 is controlled by computer control system 105to implement the CIP operation as will be discussed below. Divertervalve 445 diverts fluid to spray ball 440 in order to clean the insideof medium surge tank 410 during the CIP operation.

[0095] Filtration System

[0096]FIG. 5 illustrates prefiltration system 150 and sterile filtrationsystem 160 in further detail. Prefiltration system 150 includes aprefiltration pump 510 and a prefiltration filter 520. Prefiltrationsystem 150 receives medium product 145 from medium surge tank 140.Prefiltration pump 510 pumps medium product 145 through a non-sterileprefilter filter 520. Prefilter filter 520 is a filter membrane thatprovides variable filtration of medium product 145. Depending upon theparticular medium product 145 being prepared, the filter membrane isselected to filter particles that may range between 0.1 and 2 microns.

[0097] Medium product that has been filtered by prefiltration system 150enters sterile filtration system 160. As shown in FIG. 5, sterilefiltration system 160 operates in a clean area 180. Sterile filtrationsystem 160 includes two sterilizing filters 530A and 530B in a parallelconfiguration followed by a final sterilizing filter 540. Thisparticular configuration of sterilizing filters provides redundant 0.1or 0.2 micron filtration for medium product 145. Filtered medium product165 is output from sterile filtration system 160 and enters fill system170.

[0098] Sterilizing filters 530 and final sterilizing filter 540 aresteam sterilized via a steam in place operation which is discussed infurther detail below. In a preferred embodiment, the sterilizing filtersare steam sterilized prior to manufacturing a new batch of cell culturemedium formulation.

[0099] Fill System

[0100] As shown, fill system 170 is also contained within clean area180. Fill system 170 provides aseptic connections in clean area 180 sothat multiple medium product containers can be filled outside of cleanarea 180.

[0101] In one embodiment of present invention, fill system 170 providesa mechanism whereby multiple containers (i.e., sterile bags, carboys,glass bottles, drums, etc.) can be filled. In another embodiment of thepresent invention, fill system 170 may not be required or may bemodified. For example, an embodiment of ALMS 100 may be implemented toprovide medium product 145 directly to a bioreactor as would beapparent.

[0102] Diverter valves 505, 525 and 545 are controlled by computercontrol system 105 and used during the CIP operation as will bediscussed below. The diverter valves provide a mechanism to flushunwanted medium product through to waste disposal system 190 as well asto provide mechanisms to clean and product purge prefilter 520 andsterilizing filters 530A, 530B and 540.

[0103] ALMS Process Capability

[0104] In a preferred embodiment of the present invention, ALMS 100 isdesigned to operate with flow rates between 1,000 and 3,000 liters ormedium product per hour. Other embodiments of the present invention mayhave different flow rates depending upon the sizing and accuracy of, forexample, concentrate solution pumps 340, diluent input pump 320, andstatic mixing chamber 310.

[0105] In a preferred embodiment of the present invention, mediumproduct 165 has an intra-run homogeneity with a precision tolerance of±2.0%. Precision between production runs of medium product 165 fromidentical concentrated materials is ±3.0%. Furthermore, a pH fluctuationof medium product 165 is within ±0.1 units.

[0106] Clean In Process (CIP) and Steam In Place (SIP) ProcessOperations

[0107] ALMS 100 is designed for on-line sanitization and sterilizationin place as required. The sanitization operation is commonly referred toas “clean in place.” The sterilization operation using steam underpressure is commonly referred to as “steam in place.” A typicaloperation will require sanitization of the entire system including WFIbrake tank 230 and steam sterilization of sterile filtration system 160as well as fill system 170.

[0108] Sanitization of ALMS 100 includes the flushing of the entire ALMS100 with hot water from hot WFI 220. Hot water from hot WFI 220 isrouted through ALMS 100 via diverter valves (e.g., diverter valve 145,diverter valve 505, diverter valve 525, diverter valve 545, etc.) to andthrough spray balls (e.g., spray ball 240 and spray ball 440), andrecirculated from fill system 170 to media mixing system 130 via anappropriate conduit (shown as line 175 in FIG. 1) to flush ALMS 100. Inone embodiment of the present invention, caustic solution is added tohot water from hot WFI 220 via static mixing chamber 310 to provide ahot caustic sanitization of ALMS 100. The hot caustic is recirculated,neutralized with acid and sent to waste disposal system 190.

[0109] For sterilizing ALMS 100, steam is introduced at the sterilefiltration system 160 via a steam input port 550 located inside cleanarea 180. Steam flows through sterile filtration system 160, includingsterilizing filters 530 and final sterilizing filter 540, and fillsystem 170, and heats these components to sterilization temperatures.The temperature is monitored at appropriate points and sterilization isconfirmed using well known time/temperature parameters as would beapparent.

[0110] The by-products of the sanitization process are routed to wastedisposal system 190 as shown in various figures. In one embodiment ofthe present invention, waste disposal system treats any by-products ofALMS 100 by appropriate measures so as not to introduce any harmfulproducts into the plant's waste disposal system as would be apparent.

[0111] Computer Control System

[0112] In various embodiments of the present invention, computer controlsystem 105 is implemented using hardware, software or a combinationthereof and may be implemented in a computer system or other processingsystem. In fact, in one embodiment, the invention is directed toward acomputer system capable of carrying out the functionality describedherein. An example computer system 802 is shown in FIG. 8. Computersystem 802 includes one or more processors, such as processor 804.Processor 804 is connected to a communication bus 806. Various softwareembodiments are described in terms of this example computer system.After reading this description, it will become apparent to a personskilled in the relevant art how to implement the invention using othercomputer systems and/or computer architectures.

[0113] Computer system 802 also includes a main memory 808, preferablyrandom access memory (RAM), and may also include a secondary memory 810.Secondary memory 810 may include, for example, a hard disk drive 812and/or a removable storage drive 814, representing a floppy disk drive,a magnetic tape drive, an optical disk drive, etc. Removable storagedrive 814 reads from and/or writes to a removable storage unit 818 in awell known manner. Removable storage unit 818, represents a floppy disk,magnetic tape, optical disk, etc. which is read by and written to byremovable storage drive 814. As will be appreciated, removable storageunit 818 includes a computer usable storage medium having stored thereincomputer software and/or data.

[0114] In alternative embodiments, secondary memory 810 may includeother similar means for allowing computer programs or other instructionsto be loaded into computer system 802. Such means can include, forexample, a removable storage unit 822 and an interface 820. Examples ofsuch can include a program cartridge and cartridge interface (such asthat found in video game devices), a removable memory chip (such as anEPROM, or PROM) and associated socket, and other removable storage units822 and interfaces 820 which allow software and data to be transferredfrom the removable storage unit 818 to computer system 802.

[0115] Computer system 802 can also include a communications interface824. Communications interface 824 allows software and data to betransferred between computer system 802 and external devices. Examplesof communications interface 824 can include a modem, a network interface(such as an Ethernet card), a communications port, a PCMCIA slot andcard, etc. Software and data transferred via communications interface824 are in the form of signals which can be electronic, electromagnetic,optical or other signals capable of being received by communicationsinterface 824. Signals 826 are provided to communications interface viaa channel 828. Channel 828 carries signals 826 and can be implementedusing wire or cable, fiber optics, a phone line, a cellular phone link,an RF link and other communications channels.

[0116] In this document, the terms “computer program medium” and“computer usable medium” are used to generally refer to media such asremovable storage device 818, a hard disk installed in hard disk drive812, and signals 826. These computer program products are means forproviding software to computer system 802.

[0117] Computer programs (also called computer control logic) are storedin main memory and/or secondary memory 810. Computer programs can alsobe received via communications interface 824. Such computer programs,when executed, enable the computer system 802 to perform the features ofthe present invention as discussed herein. In particular, the computerprograms, when executed, enable processor 804 to perform the features ofthe present invention. Accordingly, such computer programs representcontrollers of the computer system 802.

[0118] In an embodiment where the invention is implement using software,the software may be stored in a computer program product and loaded intocomputer system 802 using removable storage drive 814, hard drive 812 orcommunications interface 824. The control logic (software), whenexecuted by processor 804, causes processor 804 to perform the functionsof the invention as described herein.

[0119] In another embodiment, the invention is implemented primarily inhardware using, for example, hardware components such as applicationspecific integrated circuits (ASICs). Implementation of the hardwarestate machine so as to perform the functions described herein will beapparent to persons skilled in the relevant art(s).

[0120] In yet another embodiment, the invention is implemented using acombination of both hardware and software.

[0121] Conclusion

[0122] While the invention has been particularly shown and describedwith reference to several preferred embodiments thereof, it will beunderstood by those skilled in the art that various changes in form anddetails may be made therein without departing from the spirit and scopeof the invention as defined in the appended claims.

What is claimed is:
 1. A method for continuously mixing aqueoussolutions, comprising the steps of: providing a flow controlledturbulent diluent stream; adding a plurality of chemically incompatibleconcentrate solutions to said turbulent diluent stream in such a mannerthat none of the ingredients of the concentrate solutions adverselychemically react with each other; and thereby forming a diluted mixtureof said concentrate solutions.
 2. The method of claim 1, furthercomprising the step of: adding a first of said chemically incompatibleconcentrate solutions to said turbulent diluent stream sufficiently inadvance of the addition of a second of said chemically incompatibleconcentrate solutions to prevent the ingredients of said first and saidsecond chemically incompatible concentrate solutions from adverselychemically reacting with each other.
 3. The method of claim 2, whereinsaid step of adding comprises the step of: adding said first of saidchemically incompatible concentrate solutions to said turbulent diluentstream in a manner that is approximately radially disposed from theaddition of said second of said chemically incompatible concentratesolutions to prevent the ingredients of said first and said secondchemically incompatible concentrate solutions from adversely chemicallyreacting with each other.
 4. The method of claim 2, wherein said step ofadding comprises the step of: adding said first of said chemicallyincompatible concentrate solutions to said turbulent diluent streamlinearly upstream of the addition of said second of chemicallyincompatible concentrate solutions to prevent the ingredients of saidfirst and said second chemically incompatible concentrate solutions fromadversely chemically reacting with each other.
 5. The method of claim 2,further comprising the step of: adjusting a pH level of said turbulentdiluent stream.
 6. The method of claim 5, wherein said step of adjustinga pH level comprises the step of: adding an acid solution to saidturbulent diluent stream to reduce said pH level.
 7. The method of claim5, wherein said step of adjusting a pH level comprises the step of:adding a caustic solution to said turbulent diluent stream to increasesaid pH level.
 8. The method of claim 5, wherein said step of adjustinga pH level comprises the steps of: adding an acid solution to saidturbulent diluent stream to decrease said pH level; and adding a causticsolution to said turbulent diluent stream to increase said pH level. 9.The method of claim 5, wherein said step of adjusting a pH levelcomprises the step of: adjusting a pH level of said turbulent diluentstream prior to said step of adding said plurality of chemicallyincompatible concentrate solutions to said turbulent diluent stream. 10.The method of claim 1, wherein said step of adding a plurality ofchemically incompatible concentrate solutions to said turbulent diluentstream comprises the step of: adding at least two of said plurality ofchemically incompatible concentrate solutions to said turbulent diluentstream in such a manner that none of the ingredients of said at leasttwo of said plurality of chemically incompatible concentrate solutionsadversely chemically react with each other, wherein said plurality ofchemically incompatible concentrate solutions include solutions from theset of: an acid soluble concentrate solution subgroup, a group I saltconcentrate solution subgroup, a group II salt concentrate solutionsubgroup, and a base soluble concentrate solution subgroup.
 11. Themethod of claim 3, wherein said step of adding a plurality of chemicallyincompatible concentrate solutions to said turbulent diluent streamcomprises the step of: adding a base soluble concentrate solution tosaid turbulent diluent stream in a manner that is approximately radiallydisposed to the addition of a group II salt concentrate solution so thatnone of the ingredients of said base soluble solution and said group IIsalt solution adversely chemically react with each other.
 12. The methodof claim 4, wherein said step of adding a plurality of chemicallyincompatible concentrate solutions to said turbulent diluent streamcomprises the steps of: adding an acid soluble concentrate solution tosaid turbulent diluent stream; and adding a group II salt concentratesolution to said turbulent diluent stream at a position linearlydisposed from the addition of said acid soluble concentrate solution insuch a manner that none of the ingredients of said acid solubleconcentrate solution and said group II salt concentrate solutionadversely chemically react with each other.
 13. An apparatus forcontinuously mixing aqueous solutions comprising: a static mixingchamber, having a an upstream end and a downstream end, for creating aturbulent diluent stream from a diluent introduced from the upstream endof said static mixing chamber; and a plurality of injection ports,incorporated into said static mixing chamber, for introducing aplurality of chemically incompatible concentrate solutions to saidturbulent diluent stream in such a manner that none of the ingredientsof the concentrate solutions adversely chemically react with each otherto thereby form a diluted mixture of said concentrate solutions at thedownstream end of said static mixing chamber.
 14. The apparatus of claim13, wherein said plurality of injection ports comprise a plurality ofinjection ports that are radially disposed from one another.
 15. Theapparatus of claim 13, wherein said plurality of injection portscomprise a plurality of injection ports that are linearly disposed fromone another.
 16. The apparatus of claim 13, wherein said plurality ofinjection ports includes: a first plurality of injection ports that areradially disposed from one another; and at least one injection port thatis linearly disposed from said first plurality of injection ports. 17.The apparatus of claim 13, wherein said plurality of injection portsincludes: a first plurality of injection ports incorporated into saidstatic mixing chamber and radially disposed from one another; secondplurality of injection ports incorporated into said static mixingchamber, radially disposed from one another, and linearly disposed fromsaid first plurality of injection ports; and at least one injection portincorporated into said static mixing chamber, linearly disposed fromboth said first plurality of injection ports and said second pluralityof injection ports.
 18. The apparatus of claim 16, wherein one of saidfirst plurality of injection ports introduces a base soluble concentratesolution to said turbulent diluent stream.
 19. The apparatus of claim16, wherein one of said first plurality of injection ports introduces anacid soluble concentrate solution to said turbulent diluent stream. 20.The apparatus of claim 16, wherein one of said first plurality ofinjection ports introduces a group I salts concentrate solution to saidturbulent diluent stream.
 21. The apparatus of claim 16, wherein one ofsaid first plurality of injection ports introduces a group i saltsconcentrate solution to said turbulent diluent stream.
 22. The apparatusof claim 16, wherein said at least one injection port introduces aconcentrate solution into said turbulent diluent stream.
 23. Theapparatus of claim 16, wherein said at least one injection portintroduces a solution into said turbulent diluent stream prior to saidfirst plurality of injection ports introducing any concentrate solutionsinto said turbulent diluent stream.
 24. The apparatus of claim 23,wherein said at least one injection port introduces an acid solutioninto said turbulent diluent stream.
 25. The apparatus of claim 23,wherein one of said at least one injection port introduces a causticsolution into said turbulent diluent stream.
 26. The apparatus of claim16, wherein said first plurality of injection ports do not introduceboth a group II salts concentrate solution and an acid solubleconcentrate solution into said turbulent diluent stream.
 27. Theapparatus of claim 17, wherein one of said first plurality of injectionports introduces a base soluble concentrate solution to said turbulentdiluent stream.
 28. The apparatus of claim 17, wherein one of said firstplurality of injection ports introduces an acid soluble concentratesolution to said turbulent diluent stream.
 29. The apparatus of claim17, wherein one of said first plurality of injection ports introduces agroup I salts concentrate solution to said turbulent diluent stream. 30.The apparatus of claim 17, wherein one of said first plurality ofinjection ports introduces a group II salts concentrate solution to saidturbulent diluent stream.
 31. The apparatus of claim 17, wherein said atleast one injection port introduces a solution into said turbulentdiluent stream prior to any of said first plurality of injection portsintroducing a concentrate solution into said turbulent diluent stream.32. The apparatus of claim 17, wherein said at least one injection portintroduces a concentrate solution into said turbulent diluent stream.33. The apparatus of claim 32, wherein said at least one injection portintroduces an acid solution into said turbulent diluent stream.
 34. Theapparatus of claim 32, wherein one of said at least one injection portintroduces a caustic solution into said turbulent diluent stream. 35.The apparatus of claim 17, wherein said first plurality of injectionports do not introduce both a group II salts concentrate solution and anacid soluble concentrate solution into said turbulent diluent stream.36. The apparatus of claim 17, wherein said second plurality ofinjection ports do not introduce both a group II salts concentratesolution and an acid soluble concentrate solution into said turbulentdiluent stream.
 37. The apparatus of claim 17, wherein one of said firstplurality of injection ports introduces a base soluble concentratesolution into said turbulent diluent stream and one of said secondplurality of injection ports introduces a group II salt concentratesolution into said turbulent diluent stream.
 38. A system forcontinuously preparing medium from concentrated solutions comprising: adiluent system that provides a diluent; a concentrate system thatprovides a plurality of chemically incompatible concentrate solutions;and a medium mixing system that receives said diluent, that creates aturbulent diluent stream from said diluent, and that adds said pluralityof concentrate solutions to said turbulent diluent stream in such amanner so that none of the ingredients of said plurality of concentratesolutions adversely chemically react with each other.
 39. The system ofclaim 38, wherein said medium mixing system comprises: a static mixingchamber that receives said diluent and that creates said turbulentdiluent stream from said diluent; and a plurality of injection portsincorporated into said static mixing chamber that adds said plurality ofconcentrate solutions to said turbulent diluent stream.
 40. The systemof claim 39, wherein said medium mixing system further comprises: a pumpassociated with each of said plurality of injection ports forcontrolling a flow of said plurality of concentrate solutions into saidstatic mixing chamber.
 41. The system of claim 39, wherein a first ofsaid plurality of injection ports adds a first concentrate solution intosaid static mixing chamber sufficiently in advance of a second of saidplurality of injection ports adding a second concentrate solution intosaid static mixing chamber to prevent the ingredients of said first andsecond concentrate solutions from adversely chemically reacting with oneanother.
 42. The system of claim 41, wherein said plurality of injectionports comprises: a first plurality of injection ports radially disposedfrom one another around an approximate circumference of said staticmixing chamber.
 43. The system of claim 42, wherein said plurality ofinjection ports further comprises: a second plurality of injection portsradially disposed from one another around an approximate circumferenceof said static mixing chamber and linearly disposed along a flow path ofsaid static mixing chamber.
 44. The system of claim 42, wherein saidplurality of injection ports further comprises: at least one injectionport linearly disposed along a flow path of said static mixing chamberfrom said first plurality of injections ports.
 45. The system of claim43, wherein said plurality of injection ports further comprises: atleast one injection port linearly disposed along a flow path of saidstatic mixing chamber from said first and said second plurality ofinjection ports.
 46. The system of claim 44, wherein said at least oneinjection port is linearly disposed upstream along the flow path fromsaid first plurality of injection ports.
 47. The system of claim 45,wherein said at least one injection port is linearly disposed upstreamalong the flow path from said first and said second plurality ofinjection ports.
 48. The system of claim 46, wherein said at least oneinjection port adds a solution that adjusts a pH level of said turbulentdiluent stream.
 49. The system of claim 48, wherein said at least oneinjection port adds an acid solution to said turbulent diluent stream.50. The system of claim 48, wherein said at least one injection portadds a caustic solution to said turbulent diluent stream.
 51. The systemof claim 42, wherein said first plurality of injection ports adds atleast one of the set of an acid soluble concentrate solution, a group Isalts concentrate solution, and a sodium hydroxide concentrate solution.52. The system of claim 43, wherein said first plurality of injectionports adds at least one of the set of an acid soluble concentratesolution, a group I salts concentrate solution, and a sodium hydroxideconcentrate solution.
 53. The system of claim 43, wherein said secondplurality of injection ports add at least one of the set of a group IIsalts concentrate solution and a base soluble concentrate solution. 54.A system for continuously mixing aqueous solutions, comprising: meansfor providing a flow controlled turbulent diluent stream; means foradding a plurality of chemically incompatible concentrate solutions tosaid turbulent diluent stream in such a manner that none of theingredients of the concentrate solutions adversely chemically react witheach other; and means for forming a diluted mixture of said concentratesolutions.
 55. The system of claim 54, wherein said means for addingfurther comprises: means for adding a first of said chemicallyincompatible concentrate solutions to said turbulent diluent streamsufficiently in advance of the addition of a second of said chemicallyincompatible concentrate solutions to prevent the ingredients of saidfirst and said second chemically incompatible concentrate solutions fromadversely chemically reacting with each other.
 56. The system of claim55, wherein said means for adding further comprises: means for addingsaid first of said chemically incompatible concentrate solutions to saidturbulent diluent stream in a manner that is approximately radiallydisposed from the addition of said second of said chemicallyincompatible concentrate solutions to prevent the ingredients of saidfirst and said second chemically incompatible concentrate solutions fromadversely chemically reacting with each other.
 57. The system of claim55, wherein said means for adding further comprises: means for addingsaid first of said chemically incompatible concentrate solutions to saidturbulent diluent stream linearly upstream of the addition of saidsecond of chemically incompatible concentrate solutions to prevent theingredients of said first and said second chemically incompatibleconcentrate solutions from adversely chemically reacting with eachother.
 58. The system of claim 55, further comprising: means foradjusting a pH level of said turbulent diluent stream.
 59. The system ofclaim 58, wherein said means for adjusting a pH level comprises: meansfor adding an acid solution to said turbulent diluent stream to reducesaid pH level.
 60. The system of claim 58, wherein said means foradjusting a pH level comprises: means for adding a caustic solution tosaid turbulent diluent stream to increase said pH level.
 61. The systemof claim 58, wherein said means for adjusting a pH level comprises:means for adding an acid solution to said turbulent diluent stream todecrease said pH level; and means for adding a caustic solution to saidturbulent diluent stream to increase said pH level.
 62. The system ofclaim 58, wherein said means for adjusting a pH level comprises: meansfor adjusting a pH level of said turbulent diluent stream before saidmeans for adding adds any of said plurality of chemically incompatibleconcentrate solutions to said turbulent diluent stream.
 63. The systemof claim 54, wherein said means for adding a plurality of chemicallyincompatible concentrate solutions to said turbulent diluent streamcomprises: means for adding at least two of said plurality of chemicallyincompatible concentrate solutions to said turbulent diluent stream insuch a manner that none of the ingredients of said at least two of saidplurality of chemically incompatible concentrate solutions adverselychemically react with each other, wherein said plurality of chemicallyincompatible concentrate solutions include solutions from the set of: anacid soluble concentrate solution subgroup, a group I salt concentratesolution subgroup, a group II salt concentrate solution subgroup, and abase soluble concentrate solution subgroup.
 64. The system of claim 56,wherein said means for adding a plurality of chemically incompatibleconcentrate solutions to said turbulent diluent stream comprises: meansfor adding a base soluble concentrate solution to said turbulent diluentstream in a manner that is approximately radially disposed from theaddition of a group II salt concentrate solution so that none of theingredients of said base soluble solution and said group II saltsolution adversely chemically react with each other.
 65. The system ofclaim 57, wherein said means for adding a plurality of chemicallyincompatible concentrate solutions to said turbulent diluent streamcomprises: means for adding an acid soluble concentrate solution to saidturbulent diluent stream; and means for adding a group II saltconcentrate solution to said turbulent diluent stream at a positionlinearly disposed from the addition of said acid soluble concentratesolution in such a manner that none of the ingredients of said acidsoluble concentrate solution and said group II salt concentrate solutionadversely chemically react with each other.
 66. A computer programproduct for use with a computer system, said computer program productcomprising: a computer usable medium having computer readable programcode means embodied in said medium for causing the computer system tocontrol the continuous preparation of medium from concentratedsolutions, said computer readable program code means comprising:computer readable program code means for enabling the computer system tocontrol a flow of a diluent into a static mixing chamber, wherein saidstatic mixing chamber provides a turbulent diluent stream in accordancewith said flow; computer readable program code means for enabling thecomputer system to control a flow of a plurality of chemicallyincompatible concentrate solutions into said static mixing chamber,wherein said plurality of concentrate solutions admix with saidturbulent diluent stream in such a manner that none of the ingredientsof the concentrate solutions adversely chemically react with each otherto thereby form a diluted mixture of said concentrate solutions.
 67. Thecomputer program product of claim 66, further comprising: computerreadable program code means for enabling the computer system to monitora flow of said diluted mixture out of said static mixing chamber asmeasured by a flow sensor.
 68. The computer program product of claim 67,further comprising: computer readable program code means for enablingthe computer system to adjust the flow of said diluent and the flow ofsaid plurality of concentrate solutions based on the flow of saiddiluted mixture.
 69. The computer program product of claim 66, furthercomprising: computer readable program code means for enabling thecomputer system to monitor a level of said diluted mixture in a mediumsurge tank; and computer readable program code means for enabling thecomputer system to adjust the flow of said diluted mixture into saidmedium surge tank.
 70. The computer program product of claim 69, whereinsaid computer readable program code means for enabling the computersystem to adjust the flow of said diluted mixture into said medium surgetank comprises: computer readable program code means for enabling thecomputer system to adjust the flow of said diluent into said staticmixing chamber; and computer readable program code means for enablingthe computer system to adjust the flow of said plurality of concentratesolutions into said static mixing chamber.
 71. The computer programproduct of claim 66, further comprising: computer readable program codemeans for enabling the computer system to monitor a parameter of saiddiluted mixture using a sensor located downstream from said staticmixing chamber to determine whether said diluted mixture is acceptable.72. The computer program product of claim 71, further comprising:computer readable program code means for enabling the computer system tocontrol a diverter valve that directs said diluted mixture based onwhether said diluted mixture is acceptable.
 73. The computer programproduct of claim 71, further comprising: computer readable program codemeans for enabling the computer system to monitor a pH level of saiddiluted mixture using at least one pH sensor located downstream of saidstatic mixing chamber.
 74. The computer program product of claim 73,further comprising: computer readable program code means for enablingthe computer system to adjust a pH level of said diluted mixture. 75.The computer program product of claim 74, wherein said computer readableprogram code means for enabling the computer system to adjust a pH levelof said diluted mixture comprises: computer readable program code meansfor enabling the computer system to adjust a flow of an acid solutioninto said static mixing chamber to thereby adjust the pH level of saiddiluted mixture.
 76. The computer program product of claim 74, whereinsaid computer readable program code means for enabling the computersystem to adjust a pH level of said diluted mixture comprises: computerreadable program code means for enabling the computer system to adjust aflow of a caustic solution into said static mixing chamber to therebyadjust the pH level of said diluent stream.
 77. The computer programproduct of claim 74, wherein said computer readable program code meansfor enabling the computer system to adjust a pH level of said dilutedmixture comprises: computer readable program code means for enabling thecomputer system to adjust a flow of an acid solution into said staticmixing chamber; and computer readable program code means for enablingthe computer system to adjust a flow of a caustic solution into saidstatic mixing chamber.
 78. The computer program product of claim 70,wherein said computer readable program code means for enabling thecomputer system to adjust the flow of said plurality of concentratesolutions into said static mixing chamber comprises: computer readableprogram code means for enabling the computer system to adjust a flowrate of an acid soluble concentrate solution into said static mixingchamber.
 79. The computer program product of claim 70, wherein saidcomputer readable program code means for enabling the computer system toadjust the flow of said plurality of concentrate solutions into saidstatic mixing chamber comprises: computer readable program code meansfor enabling the computer system to adjust a flow of a group I saltconcentrate solution into said static mixing chamber.
 80. The computerprogram product of claim 70, wherein said computer readable program codemeans for enabling the computer system to adjust the flow of saidplurality of concentrate solutions into said static mixing chambercomprises: computer readable program code means for enabling thecomputer system to adjust a flow of a group II salt concentrate solutioninto said static mixing chamber.
 81. The computer program product ofclaim 70, wherein said computer readable program code means for enablingthe computer system to adjust the flow of said plurality of concentratesolutions into said static mixing chamber comprises: computer readableprogram code means for enabling the computer system to adjust a flow ofa base soluble concentrate solution into said static mixing chamber. 82.A medium product produced by the method of claim
 1. 83. The mediumproduct of claim 82, wherein said medium product is selected from thegroup consisting of a 1× cell culture medium, a concentrated cellculture medium, and a buffered salt solution.